Wash-free instant detection of giant plasma membrane vesicles
Wash-free instant detection of giant plasma membrane vesicles
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免洗即时检测巨型质膜囊泡
DOI：
10.1016/j.ab.2018.07.012
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发表时间：
2018
影响因子：
2.9
通讯作者：
Saitoh Hisato
中科院分区：
文献类型：
--
作者：
Okada Saya;Yankawa Satoshi;Saitoh Hisato
巨型质膜囊泡（GPMV）是细胞暴露于多聚甲醛或其他应激下产生的大型细胞外囊泡，为阐明细胞表面动力学提供了一个实验系统。在这里，我们表明，将膜渗透性荧光 RNA 指示剂吖啶橙和硫黄素 T 添加到由培养的 HeLa 细胞制备的含有 GPMV 的溶液中，足以实现几乎所有 GPMV 的荧光可视化。我们的研究结果提供了一种使用非脂质型荧光染料进行 GPMV 检测的免洗即时方法，这对于有兴趣使用 GPMV 研究细胞膜动力学和生物化学的研究人员来说应该很有用。
Giant plasma membrane vesicles (GPMVs) are large extracellular vesicles produced by the exposure of cells to paraformaldehyde or other stresses, providing an experimental system to elucidate cell surface dynamics. Here we show that addition of the membrane permeable fluorescent RNA-indicators, acridine orange and thioflavin T, to GPMV-containing solutions prepared from cultured HeLa cells was sufficient for the fluorescent visualization of seemingly all GPMVs. Our findings provide a wash-free instant method using non-lipid-type fluorescent dyes for GPMV detection, which should be useful for researchers interested in studying cell membrane dynamics and biochemistry using GPMVs.